摘要
目的 建立选择性培养牛视网膜血管内皮细胞和周细胞的简要方法。方法 采用机械除杂法或等密度沉降分离法 ,培养皿用不同的基质成分包埋 ,培养液中加入有利于细胞生长的添加物。结果 原代培养的牛视网膜血管内皮细胞和周细胞纯净度 >95 % ,能连续传代。周细胞在传代后不规则形状及细胞内微丝更明显。视网膜血管内皮细胞融合后呈“铺路石”样改变。视网膜血管内皮细胞对Ⅷ因子相关抗体染色阳性。视网膜周细胞对单克隆抗体α 平滑肌肌动蛋白抗体染色阳性。结论 用明胶包被培养皿 ,在培养液中加入肝素、内皮细胞生长因子及高浓度的胎牛血清可获得较纯的内皮细胞。而周细胞最适合的培养液为DMEM加入 2 0 g·L-1胎牛血清。
Objective To establish simple methods for selective culture of bovine retinal endothelial cell and pericytes.Methods Cell weeding or seperation were used ,in addition,culture dishes were coated with different matrix components and supplements benefit to cell growth were added into culture media.Results Relative pure bovine retinal vascular endothelial cells and pericytes were obtained selectively (purity>95%)and could be passaged serially.Passaged pericytes showed higyly irregular peripheries and more silky structure in cells, and positive staining for a monoclonal antibody α smooth muscle actin.Retinal endothelial cells showed ‘stone’ type when confluent,and positive staining for factor Ⅷ antibody.Conclusion The suitable condition for endothelial cell were use of gelatin coat culture dishes, and adding of heparin,endothelial growth factor and highly fetal serum into culture media. The suitable condition for pericytes are adding 20 g·L -1 fetal serum into DMEM. Percoll can also be used to selective culture of endothelial cells and pericytes.
出处
《眼科新进展》
CAS
2004年第1期22-25,共4页
Recent Advances in Ophthalmology
